Sensitivity of the HEp-2000 substrate for the detection of anti-SSA/Ro60 antibodies

Clin Rheumatol. 2000;19(4):291-5. doi: 10.1007/s100670070048.


Anti-SSA/Ro antibodies are the most prevalent type of antinuclear antibody (ANA). Anti-SSA/ Ro-positive sera may recognise two proteins: a 52 kDa (Ro52) and a 60 kDa (Ro60) subunit. We studied the sensitivity for Ro60 detection using the HEp-2000 substrate, which consists of HEp-2 cells transfected with Ro60 cDNA in an anti-SSA/Ro-positive population consecutively identified by double immunodiffusion (DID) with thymus/spleen nuclear extract and line immunoassay (LIA) with recombinant Ro52 and Ro60. One hundred and twenty-seven consecutive anti-SSA/Ro-positive sera defined by DID with thymus/spleen nuclear extract and LIA using recombinant Ro52 and Ro60 were analysed on HEp-2000 and DID with natural Ro60. Of these, 91 were anti-Ro60 positive on LIA and/ or DID with natural Ro60. The HEp-2000 substrate detected 70/91 (sensitivity 77%) and correlated strongest with DID. Most of the missed anti-Ro60-positive sera had high ANA intensity. The substrate did not detect monospecific anti-Ro52 antibodies (sensitivity 9.7%; 3/31). HEp-2000 substrate can therefore be considered a reliable, simple and alternative method for DID in the detection of anti-Ro60 reactivity. Special follow-up should be given to sera with strong ANA patterns in which the SSA/Ro60 staining pattern may be hidden.

Publication types

  • Comparative Study

MeSH terms

  • Adult
  • Antibodies, Antinuclear / blood*
  • Antigens, Nuclear
  • Autoantigens / immunology*
  • DNA-Binding Proteins / immunology
  • Female
  • Fluorescent Antibody Technique, Indirect
  • Humans
  • Immunoassay
  • Immunodiffusion
  • Infant, Newborn
  • Lupus Erythematosus, Systemic / immunology
  • Male
  • Nuclear Proteins / immunology*
  • Ribonucleoproteins / immunology*
  • Sensitivity and Specificity
  • Staining and Labeling
  • Transfection
  • Tumor Cells, Cultured


  • Antibodies, Antinuclear
  • Antigens, Nuclear
  • Autoantigens
  • DNA-Binding Proteins
  • Nuclear Proteins
  • Ribonucleoproteins